The binding of precipitant ions in the tetragonal crystals of hen egg white lysozyme

Abstract

The bonds between lysozyme molecules and precipitant ions in single crystals grown with chlorides of several metals are analysed on the basis of crystal structure data. Crystals of tetragonal hen egg lysozyme (HEWL) were grown with chlorides of several alkali and transition metals (LiCl, NaCl, KCl, NiCl₂ and CuCl₂) as precipitants and the three-dimensional structures were determined at 1.35?Å resolution by X-ray diffraction method. The positions of metal and chloride ions attached to the protein were located, divided into three groups and analysed. Some of them, in accordance with the recently proposed and experimentally confirmed crystal growth model, provide connections in protein dimers and octamers that are precursor clusters in the crystallization lysozyme solution. The first group, including Cu⁺², Ni⁺² and Na⁺¹ cations, binds specifically to the protein molecule. The second group consists of metal and chloride ions bound inside the dimers and octamers. The third group of ions can participate in connections between the octamers that are suggested as building units during the crystal growth. The arrangement of chloride and metal ions associated with lysozyme molecule at all stages of the crystallization solution formation and crystal growth is discussed.

Communicated by Ramaswamy H. Sarma     

Peculiarities of functional activity of circulating phagocytes in patients with different forms of drug-resistant pulmonary tuberculosis

Functional activity of circulating phagocytes (macrophages and neutrophils — Ns) was studied in 30 patients with infiltrative and 30 patients with fibro-cavernous pulmonary tuberculosis (IPT and FCPT, respectively), characterized by similar biological properties of mycobacterium tuberculosis (MTB), dissemination of the process, and manifestations of intoxication. Differences of the functional activity of both types of cells depending on the PT form were found: a more significant increase in the oxygen-depending activity in FCPT while bactericide potential estimated with the zymosan induced NST-test was more pronounced in IPT patients. These data correlate with the blood levels of neopterin and elastase, the markers of the mononuclear and neutrophil activity, respectively. Involvement of adenosine deaminase (ADA) and neopterin in realization of intracellular oxygen-dependent processes was demonstrated. Results of the multivariate analysis of the whole set of the studied phagocyte parameters, reflecting their different roles in this pathological process demonstrated a prevailing role of mononuclears in newly diagnosed IPT and neutrophils in the chronic progressive process.     

Changes in the Balance between Membrane-Bound and Soluble Forms of CD95 (Fas) during Selection of Tumor Cells in vivo

Studies concerning the expression of the receptor CD95 (Fas) by tumor cells and the role of this protein in apoptosis induced by the effector host cells that bear Fas-ligand are mainly focused on the membrane-bound form of Fas. There are only a few data about the production of the soluble form of Fas by the tumor cells, its role in the interaction with the effector host cells, and the possible changes in the synthesis of this protein during tumor progression. In the present work, three in vivo transformed parental cell lines of different origin and 24 of their variants isolated after a short cycle of natural selection in vivo were studied. It was demonstrated for the first time that: 1) production of the soluble Fas by all selected in vivo variant tumor cell lines increased significantly (2-10-fold) in comparison to the initial (parental) cell lines and did not depend on the origin of the parental lines. At the same time, the expression of the membrane bound form of Fas decreased considerably; 2) variations of the balance between membrane-bound and soluble forms of Fas in selected in vivo variant cells and the expression of the [H₂O₂ ^CA + PGE^S]-phenotype by these cells (this phenotype determines one of the essential mechanisms of the protection of a tumor cell in vivo) possibly represent independent secondary changes acquired during tumor progression in vivo.     

Effect of Forced Treadmill Running on Skeletal Muscle Myokine Levels in Mice with a Model of Type II Diabetes Mellitus

Journal of Evolutionary Biochemistry and Physiology - The effect of forced treadmill running on the level of some cytokines in skeletal muscles of mice with a model of type II diabetes mellitus was...     

Pre-crystallization phase formation of thermolysin hexamers in solution close to crystallization conditions

Communicated by Ramaswamy H. Sarma     

Intracellular Localization and the Mechanisms of Photodynamic Action of 131-[2-(Guanidinyl)ethylamino] Chlorin e6 Dimethyl Ester

Russian Journal of Bioorganic Chemistry - It has been found that the photosensitizer 131-[2-(guanidinyl)ethylamino] chlorin e6 dimethyl ester accumulates predominantly in lysosomes, partially in...     

Functional activity of phagocytes of bronchoalveolar lavage in patients with fibro-cavernous and infiltrative pulmonary tuberculosis

Functional activity of the bronchoalveolar lavage fluid (BALF) phagocytes was studied in 33 and 16 patients with fibro-cavernous and infiltrative pulmonary tuberculosis (FCPT and IPT, respectively). Complex examination of BALF, alveolar macrophages and neutrophils sedimented from BALF has shown interrelationship between functional activity of the cells and the form of pulmonary tuberculosis. Higher neopterin content and activity of elastase mainly secreted into BALF by activated alveolar macrophages and neutrophils, respectively, reflect higher secretory activity of both types of cells in FCPT. In FCPT this is combined with higher bactericidal activity of neutrophils, which significantly correlates with their adenosine deaminase (ADA) activity. Comparison of changes of the biochemical parameters studied in BALF (neopterin, elastase, ADA and its isoenzymes, 2-deoxy-ADA) and bactericidal activity of the sedimented cells obviously reflects different sides of BALF phagocytes functioning. Taking into consideration modern concepts on the mechanisms of regulation of phagocyte cells one may suggest the existence of differences in intercellular interactions in various forms of pulmonary tuberculosis.     

Alternative production of Bcl-2 and Bax by tumor cells determines the rates of in vivo tumor progression: suggested mechanisms

The hypothesis tested in the study suggests that mechanisms of the earlier described delayed or accelerated tumor progression may be regulated by the antiapoptotic and proapoptotic cellular programs activated in stress reactions of transformed cells to the host normal cellular environment. Therefore, spontaneously transformed hamster cell line STHE, its bcl-2-transduced line STHE-Bcl-2, and 64 of their descendant tumor cell variants naturally selected in two in vivo regimes (local tumor growth versus dissemination) were examined. The role of Bcl-2 and the possible activation of endogenous death-signaling Bax, Ras, and HSP90/HSP70 stress proteins in STHE (Bcl-2+/-) tumor cell variants were studied in dynamics of in vivo tumor progression. The data demonstrate: (1) Immediate in vivo activation of Bax and of HSP90/HSP70 stress proteins in disseminated STHE cells on the background of accelerated tumor progression; (2) No immediate activation of Bax and the gradual downregulation of Bcl-2 in STHE-Bcl-2 cells on the background of delayed tumor progression; (3) Alternative and mutually suppressive character of Bcl-2 and Bax expression in both regimes of tumor progression; (4) In the later stages of tumor progression, the regular transit of the initial Bcl-2 antiapoptotic, Bax-suppressing program, and the delayed tumor progression towards Bcl-2 loss, activation of Bax, and acceleration of tumor progression. Thus, the delay of tumor progression is apparently determined by the ability of Bcl-2-expressing tumor cells to extinguish the cell-damaging environmental stress signals and Bax activation, while its acceleration correlates with Bcl-2 loss, activation of proapoptotic Bax, and tumor cells damage.